Biopharmaceutical characterization is required throughout all stages of the drug pipeline, from discovery, through development, production, and quality control. Liquid chromatography (LC) with high-resolution accurate-mass mass spectrometry (MS) approaches are used to structurally characterize therapeutic proteins—such as monoclonal antibodies (mAbs)—to assess purity and heterogeneity characteristics including quantification of the various glycoforms and protein variants. New, rapid, and simplified methods coupling ultra high performance chromatography (UHPLC) with Orbitrap-based mass spectrometers give fast, confident insight into the proteoform profile for direct comparison of biotherapeutic products.

This two-part webcast series, featuring experts from Genentech and Thermo Fisher Scientific, will begin with characterization of intact proteins using denaturing and native conditions in LC-MS analysis with Orbitrap mass spectrometers. Specific applications to be discussed include common challenges such as identifying low abundance isoforms in simple antibodies, resolving heterogeneity in antibody drug conjugates (ADCs) without sample treatment, assessing purity of bispecific antibodies, and measuring protein-ligand affinities.

The second webcast will feature sample preparation considerations for LC-MS analysis of antibodies on the intact and subunit level, both performed under denaturing conditions. Experts will show the workflow for intact mass analysis and discuss chromatographic options for fast desalting to enable high-throughput analysis. Other topics include the entire workflow for subunit analysis consisting of optimized sample preparation and details to keep an eye on; high-resolution chromatographic separation; mass analysis on a quadrupole-Orbitrap mass spectrometer; and data analysis.